Figure 4.

Disruption of spindles and spindle fiber formation after introduction of Nek2 siRNA or ASO. Immunofluorescence image of ASO or siRNA transfected (A) M231 and (B) M468 breast cancer cells. Both cell lines were transfected with Nek2 siRNA or ASO before cell cycle synchronization with thymidine. Synchronized cells were either untreated or transfected with Nek2 100 nM siRNA or 50 nM ASO, fixed in cold methanol, and stained with antibodies against γ-tubulin (gray image and red spot in merge) and α-tubulin (green). DNA was counterstained with DAPI (blue) (magnification; bar, 50 μm). Both Nek2 depleted cell lines demonstrated diffused spindle poles, unusual microtubule formation, weak α-tubulin staining, and misaligned chromosomes. The results are representative of 3 independent trials.